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Preparation method and application of double-ligand-carrying interleukin ultrasonic targeting microbubbles

A technology targeting microbubbles and interleukins, which is applied in the field of biomedicine to achieve good imaging characteristics, improved sensitivity and specificity, and strong targeting adhesion performance

Inactive Publication Date: 2020-12-01
陆永萍
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is a lack of a targeted ultrasound contrast agent and its preparation method that can specifically / effectively assess the inflammatory response of cardiovascular diseases

Method used

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  • Preparation method and application of double-ligand-carrying interleukin ultrasonic targeting microbubbles
  • Preparation method and application of double-ligand-carrying interleukin ultrasonic targeting microbubbles
  • Preparation method and application of double-ligand-carrying interleukin ultrasonic targeting microbubbles

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method for preparing a dual-ligand interleukin-carrying ultrasound-targeted microbubble of the present invention includes the following steps: (1) Flow cytometry detects different doses of biotinylated IL-6 monoclonal antibody and IL-8 monoclonal antibody and Binding rate of TargestarSA microbubbles:

[0034] Mix different doses of biotin and fluorescent FITC-labeled IL-6 monoclonal antibodies, biotin and fluorescent Rhodamine-labeled IL-8 monoclonal antibodies, and 400ul Targestar SA microbubbles, respectively, and react at room temperature for 30 minutes. Fully combined, sampled, pressed into tablets, observed under a fluorescence microscope, and the binding rate of antibodies and microbubbles was detected by flow cytometry;

[0035] The binding rate of antibodies to microvesicles was detected by flow cytometry and grouped as follows:

[0036] G0: naked microbubble group (400ul Targestar SA microbubbles),

[0037] G1: 5ug IL-6-G antibody, 5ug IL-8-R antibody and 4...

Embodiment 2

[0044]A method for preparing a dual-ligand interleukin-carrying ultrasound-targeted microbubble of the present invention includes the following steps: (1) Flow cytometry detects different doses of biotinylated IL-6 monoclonal antibody and IL-8 monoclonal antibody and Binding rate of TargestarSA microbubbles:

[0045] Mix different doses of biotin and fluorescent FITC-labeled IL-6 monoclonal antibodies with biotin and fluorescent Rhodamine-labeled IL-8 monoclonal antibodies and 400ul Targestar SA microbubbles, respectively, and react at room temperature for 20 minutes, with constant shaking during the period to ensure that the antibodies and microbubbles Fully combined, sampled, pressed into tablets, observed under a fluorescence microscope, and the binding rate of antibodies and microbubbles was detected by flow cytometry;

[0046] The binding rate of antibodies to microvesicles was detected by flow cytometry and grouped as follows:

[0047] G0: naked microbubble group (400ul...

Embodiment 3

[0050] A method for preparing a dual-ligand interleukin-carrying ultrasound-targeted microbubble of the present invention comprises the following steps: in step (1), flow cytometry detects different doses of biotinylated IL-6 monoclonal antibody and IL-6 The binding rate of 8 mAbs to TargetstarSA microbubbles:

[0051] Mix different doses of biotin and fluorescent FITC-labeled IL-6 monoclonal antibodies with biotin and fluorescent Rhodamine-labeled IL-8 monoclonal antibodies and 400ul Targestar SA microbubbles, respectively, and react at room temperature for 40 minutes. Fully combined, sampled, pressed into tablets, observed under a fluorescence microscope, and the binding rate of antibodies and microbubbles was detected by flow cytometry;

[0052] (2) Preparation of ultrasonic microbubbles carrying IL-6 and IL-8 double antibodies:

[0053] Based on the result of the flow cytometry detection of the binding rate in step (1), the microbubbles with the highest binding rate were ...

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Abstract

The invention discloses a preparation method and application of double-ligand-carrying interleukin ultrasonic targeting microbubbles. The preparation method of the double-ligand-carrying interleukin ultrasonic targeting microbubbles comprises the following steps: (1) detecting the binding rates of biotinylated IL-6 monoclonal antibodies and IL-8 monoclonal antibodies with different doses and TargestarSA microbubbles by virtue of a flow cytometry; (2) preparing ultrasonic microbubbles carrying double antibodies of IL-6 and IL-8: adding 100 [mu]g of an IL-6 antibody labeled by biotin and fluorescence (FITC) and 100 [mu]g of an IL-8 antibody labeled by biotin and fluorescence (Rhodamin) into 1 ml of streptavidin-coated Targestar SA microbubble in a vial by using a trace sampling gun, performing incubating at room temperature until the freeze-dried powder is not completely dispersed, so as to obtain a uniform white emulsion suspension, namely, to obtain the ultrasonic targeting microbubbles carrying IL-6 and IL-8 double antibodies.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a preparation method and application of a dual-ligand interleukin-carrying ultrasonic targeting microbubble. Background technique [0002] Ultrasound contrast agent is a red blood cell tracer that can pass through the capillary network to generate an ultrasound signal in the myocardium that is proportional to the blood flow and enhance the echo in the lesion area. In recent years, with the rise of ultrasonic molecular probe technology, targeted ultrasonic microbubbles have become one of the hot spots in current ultrasonic diagnostic research. As a molecular imaging agent, specific antibodies or ligands can be linked to particles smaller than The surface of the ultrasound contrast agent of red blood cells and specifically binds to it. After peripheral intravenous injection, it accumulates in the target tissue specifically with the blood circulation, enhances the echo signal o...

Claims

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Application Information

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IPC IPC(8): A61K49/22
CPCA61K49/221A61K49/223
Inventor 陆永萍孙月杨寒凝徐丽荣王禹雪李云燕陈建福张静秋汤跃跃
Owner 陆永萍
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